The regulatory role of natural progesterone in barrier immunity

摘要：孕酮(P4)是一种调节月经生理变化的关键性激素 周而复始。我们和其他人已经证明，它对局部的免疫系统也有抗炎作用。 女性生殖道(FRT)，但其机制尚不清楚。也有迹象表明， 它的免疫效果超越了FRT，并从系统上影响免疫。P4的S效果很可能被 雌二醇(E2)的促炎作用越强。对背后机制的详细了解 P4‘S在局部和全身部位的免疫活性及其与E2的相互作用可能有助于 通过解释女性健康的性别和月经周期相关的炎症性波动 疾病。一些炎症性疾病在月经周期中波动，这表明P4 全身性免疫效应。例如，哮喘和炎症性肠病在P4-Low恶化 滤泡期，而类风湿性关节炎通常在P4主导的黄体期改善。另一方面， P4诱导的免疫抑制可限制对疫苗接种的反应或对感染的抵抗力。 P4在月经周期的黄体期激增，达到卵泡期水平的40倍。我们的 数据显示，FRT在不同时期有明显的免疫差异：黄体对巨噬细胞的抑制- P4的热带趋化因子。单核/巨噬细胞和树突状细胞(DC)是抗原提呈细胞 (APC)，为驻留记忆T细胞(TRM)的定位和存活提供关键信号。 我们建议补充人类和小鼠的研究：(1)具有APC水平研究的人类队列， 探索它们在FRT和其他屏障部位与TRM的联系；(2)小鼠实验证实 P4‘S结合E_2的免疫学特性及验证阴道CCl_2/CCl_4的假说 给药部分逆转P4诱导的免疫抑制。 在目标1中，我们将招募一个临床队列来研究人类以P4为主的黄体期的影响。 APC和TRM在FRT、胃肠道、上呼吸道、小鼠的激活和功能 皮肤，还有血。我们将使用基于MSD的免疫因子分泌物分析和流式细胞术 黏膜细胞的单细胞RNA测序解决三个假说：(1)抑制嗜酸性磷酸酶 趋化因子的产生是黄体期屏障部位免疫抑制的标志；(2)粘膜 APC在黄体期比卵泡期炎症更少或更少；以及(3)APC和细胞因子 黄体期的变化与TRM数量和/或激活状态减少有关。 在目标2中，我们将使用小鼠模型来建立P4治疗、趋化因子之间的因果联系 生产，以及APC和TRM频率和激活状态。我们将提出两个假设：(1)P4 抑制主要屏障组织中CCL2和CCL4的产生，降低APC和TRM频率和/或 (2)经阴道给予CCL2/4可阻止P4对APC和TRM的抑制。

ABSTRACT: Progesterone (P4) is a key sex hormone governing the physiological changes of the menstrual cycle. We and others have shown that it also has anti-inflammatory effects on the immune system locally in the female reproductive tract (FRT), but the mechanisms are not well understood. There are also indications that its immune effects reach beyond the FRT and affect immunity systemically. P4’s effects are likely modified by the more pro-inflammatory effects of estradiol (E2). A detailed understanding of the mechanisms behind P4’s immune activities at local and systemic sites, and its interactions with E2, may contribute to women’s health by explaining sex-based and menstrual cycle-related fluctuations in inflammatory diseases. Some inflammatory diseases fluctuate during the menstrual cycle, suggesting that P4 has systemic immune effects. For example, asthma and inflammatory bowel disease worsen in the P4-low follicular phase, while rheumatoid arthritis often improves in the P4-dominant luteal phase. On the other hand, P4-induced immunosuppression could limit the response to vaccination or resistance to infection. P4 surges during the luteal phase of the menstrual cycle, reaching >40 times above follicular phase levels. Our data show a clear immunological difference in the FRT between the phases: luteal inhibition of macrophage- tropic chemokines by P4. Monocytes/macrophages and dendritic cells (DC) are antigen-presenting cells (APC), which provide key signals for the localization and survival of resident memory T cells (TRM). We propose complementary human and mouse studies: (1) a human cohort with studies at the APC level, probing their connections to TRM in the FRT and other barrier sites and (2) mouse experiments to corroborate P4’s immunological properties in combination with E2 and to test the hypothesis that vaginal CCL2/CCL4 administration partially reverses P4-induced immune suppression. In Aim 1, we will recruit a clinical cohort to study the effects of the P4-dominant luteal phase in humans on APC and TRM activation and function in the FRT, the gastrointestinal tract, the upper respiratory tract, the skin, and the blood. We will use MSD-based immune factor profiling of secretions and flow cytometry and single cell RNA sequencing of mucosal cells to address three hypotheses: (1) inhibition of APC-tropic chemokine production is a hallmark of barrier site immunosuppression during the luteal phase; (2) mucosal APCs are fewer or less inflammatory during the luteal than the follicular phase; and (3) the APC and cytokine changes during the luteal phase associate with lower number and/or activation status of TRM. In Aim 2, we will use mouse models to establish a causal link between P4 treatment, chemokine production, and APC & TRM frequencies and activation state. We will address two hypotheses: (1) P4 inhibits production of CCL2 and CCL4 in major barrier tissues, reducing APC and TRM frequencies and/or functional potential; and (2) vaginal CCL2/4 administration prevents P4-induced suppression of APC and TRM.